DETERMINAÇÃO DA MELHOR CONDIÇÃO DE CULTIVO PARA OBTENÇÃO DE UM EXTRATO RICO EM FLAVONOIDES DAS FOLHAS DE Moringa oleifera Lam.: ESTUDO FITOQUÍMICO, DE TOXICIDADE E DE ATIVIDADE ANTI-INFLAMATÓRIA in vitro.
Moringa oleifera; Cultivation; Flavonoids; Quality control; Inflammation; Safety;
Moringa oleifera Lam. (Moringaceae), popularly known as moringa or white acacia, native to
India, is currently cultivated in several countries around the world, and distributed in many
regions from Brazil. It is a fast growing perennial plant, with significant content of nutrients
such as vitamins, minerals, amino acids, and proteins. All parts of the plant are utilized (seeds,
leaves, fruits, branches and roots) medicinally, ornamentally, industrially or for food for human
and animal use. It can also be an alternative source for the improvement of drinking water
quality and for fertilization in the cultivation of other plant species. However, when it comes to
therapeutic use, the leaves are the most used part of the plant, mainly to prevent and treat
diabetes. However, the problem regarding the therapeutic use of M. oleifera is that there is no
definition about the chemical composition content to ensure the safe and effective use of
derivatives of the species. Within this context, the present work aimed to standardize the culture
conditions and optimize the extraction conditions to obtain a flavonoid-rich extract from M.
oleifera leaves, evaluate the in vitro and in vivo toxicity and the in vitro anti-inflammatory
activity. It was also part of the thesis to compare different samples of moringa commercialized
in Brazil and the United States. For this, initially the study of different soil cultivation
conditions (mineral fertilization, mineral fertilization + organic compost, mineral fertilization
+ charcoal, and the control, soil without fertilization) and two harvesting seasons (dry and rainy)
of the leaves of the species M. oleifera was developed, aiming to identify the best condition to
obtain the highest content of flavonoids in the leaves. The extraction process was then
optimized to concentrate the flavonoids. This optimized extraction method was applied to
samples from different growing and harvesting conditions to select the most promising sample.
All extracts were submitted to analysis of total content of phenols, flavonoids, sugars and
proteins, yield analysis and chromatographic profile by ultra efficient liquid chromatography
coupled to mass spectrometer (CLUE-EM/MS). The extract from the condition of cultivation
with mineral fertilization was the one with the highest content of phenols, flavonoids, proteins
and good yield, being selected for the next steps of the study. In the analysis by CLUE-EM/MS,
it was possible to characterize eith majority substances predominantly from the flavonoid class,
with prevalence of flavonol type nuclei. Cytotoxicity and oral toxicity (acute and repeated
doses) of the optimized flavonoid-rich extract were evaluated. And was observed safety for
acute use until a dose of 2,000mg/kg and subchronic use at a dose of 400 mg/kg. Finally, a
comparative analysis was made of the extracts of all culture conditions against commercial
samples that were purchased in Brazil and the United States, and a comparison between plant
drug (pulverized dried leaves) and plant derivative (plant extract). A visual comparison was
made, of chemical profile by CLAE-DAD and by in vitro biological activity, antioxidant and
anti-inflammatory, with two macrophage cell lines. The results showed great variation visually
and between the phytochemical profiles of the commercial samples, regardless of the country
of their commercialization. And the phytochemical and in vitro activity evaluation showed
better results for the vegetal extract samples, compared to the vegetal drugs. This set of results
contributes to the quality control of the optimized extract of M. oleifera leaves, to the future
characterization of the species' chemical markers and to the development of a with standardized
process to obtain products with quality, efficacy and safety for the population.